Endo S |NEB酶试剂 New England Biolabs

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产品信息

Endo S |
 
Endo S is an endoglycosidase with a uniquely high specificity for removing N-linked glycans from the chitobiose core of the heavy chain of native IgG (1). Endo S is tagged with a chitin binding domain (CBD) for easy removal from a reaction and is supplied glycerol free for optimal performance in HPLC and MS intensive methods.

产品来源

Endo S is cloned from Streptococcus pyogenes and overexpressed as a fusion to the chitin binding domain in E. coli.

特异性

Detailed Specificity
X = protein, peptide, Asparagine, or free glycan, as Endo S does not have a strict peptide requirement for activity. Endo S is active on a substrate with or without core α(1-6)fucosylation as well as a with or without a bisecting N-acetylglucosamine. Triantennary and tetrantennary sialyated or asialo glycans are not a substrate for Endo S (1).

产品类别:
Endoglycosidases Products,
Proteome Analysis Products

应用:
Expression Systems,
Glycan Sequencing,
Proteomics,

Recombinant Glycoprotein Expression,

Glycoprotein Analysis

  • 产品组分信息

    本产品提供以下试剂或组分：

    NEB # 名称 组分货号 储存温度 数量 浓度
    • P0741S     4    
        Endo S P0741SVIAL 4 1 x 0.03 ml 200,000 units/ml
        GlycoBuffer 1 B1727SVIAL -20 1 x 1 ml 10 X
    • P0741L     4    
        Endo S P0741LVIAL 4 1 x 0.15 ml 200,000 units/ml
        GlycoBuffer 1 B1727SVIAL -20 1 x 1 ml 10 X

  • 特性和用法

    单位定义

    One unit is defined as the amount of enzyme required to remove > 95% of the carbohydrate from 5 μg of native mouse monoclonal IgG in 1 hour at 37°C in a total reaction volume of 10 µl.

    反应条件

    1X GlycoBuffer 1
    Incubate at 37°C

    1X GlycoBuffer 1
    5 mM CaCl2
    50 mM sodium acetate
    (pH 5.5 @ 25°C)

    贮存溶液

    20 mM Tris-HCl
    50 mM NaCl
    5 mM EDTA
    pH 7.5 @ 25°C

    热失活

    55°C for 10 minutes

    分子量

    实际: 136000 daltons

    单位活性检测条件

    5 µg of IgG in 1X GlycoBuffer 1 are incubated with two-fold dilutions of Endo S for 1 hour at 37°C. Separation of reaction products is visualized by SDS-PAGE.

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  • 注意事项

    1. Recommended storage temperature is 4°C,avoid repeat freeze-thaw cycles.
    2. Removal of Endo S from thedeglycosylation reaction can be scaled uplinearly with larger volumes of chitin magneticbeads.
    3. The binding capacity of Chitin Magnetic Beads is 1 mg of CBD-tagged protein / mL of beads.

  • 参考文献

    1. Bielik, A., McLeod, E. and Magnelli, P., New England Biolabs,Inc., unpublished results.. Unpublished observation

操作说明、说明书 & 用法

  • 操作说明

    1. Reaction Conditions for Endo S (P0741)
    2. Endo S Removal Magnetic Chitin Bead Protocol (P0741)

  • 应用实例

    • AppNote_Glycan_Analysis_of_Murine_IgG_by_Enzymatic_Digestion_with_Endo_S_and_PNGase_F
    • Glycan Analysis of Murine IgG by Enzymatic Digestion with Endo S and PNGase F Followed by Mass Spectrometric Analysis

工具 & 资源

  • 选择指南

    • Endoglycosidase Selection Chart

FAQs & 问题解决指南

  • FAQs

    1. Is Endo S tagged?
    2. What is the difference between PNGase F and Endo S?
    3. How much Endo S should I use to deglycosylate a glycoprotein under native conditions?
    4. What is the preferred substrate for Endo S?
    5. What are the typical reaction conditions for Endo S?
    6. How do I eliminate Endo S from a reaction?
    7. What is the binding capacity of the Magnetic Chitin Beads used to remove Endo S?
    8. Is Endo S compatible with downstream analysis such as HPLC and Mass Spectrometry?
    9. Why have the NEB Glycosidase enzymes changed reaction buffers? What are the new reaction buffers and can I still use an enzyme with its old buffer? Where can I find the composition of the old buffers?
    10. What are Glycosidases and their uses?